Influence of in Vitro Assay Setup on the Apparent Cytotoxic Potency of Benzalkonium Chlorides
| Autor: | Groothuis, Floris A., Timmer, Niels, Opsahl, Eystein, Nicol, Beate, Droge, Steven T.J., Blaauboer, Bas J., Kramer, Nynke I., LS IRAS Tox CMT/ATX, dIRAS RA-1, One Health Toxicologie |
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| Přispěvatelé: | LS IRAS Tox CMT/ATX, dIRAS RA-1, One Health Toxicologie |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cell Survival
010501 environmental sciences Toxicology 01 natural sciences 03 medical and health sciences Structure-Activity Relationship Structure–activity relationship Animals Bovine serum albumin Alkyl Cells Cultured 030304 developmental biology 0105 earth and related environmental sciences EC50 chemistry.chemical_classification 0303 health sciences Chromatography biology Dose-Response Relationship Drug Molecular Structure Sorption General Medicine In vitro Dose–response relationship chemistry Oncorhynchus mykiss biology.protein Benzalkonium Compounds Fetal bovine serum |
| Zdroj: | Chemical Research in Toxicology, 32(6), 1103. American Chemical Society |
| ISSN: | 0893-228X |
| Popis: | [Image: see text] The nominal concentration is generally used to express concentration–effect relationships in in vitro toxicity assays. However, the nominal concentration does not necessarily represent the exposure concentration responsible for the observed effect. Surfactants accumulate at interphases and likely sorb to in vitro system components such as serum protein and well plate plastic. The extent of sorption and the consequences of this sorption on in vitro readouts is largely unknown for these chemicals. The aim of this study was to demonstrate the effect of sorption to in vitro components on the observed cytotoxic potency of benzalkonium chlorides (BAC) varying in alkyl chain length (6–18 carbon atoms, C(6–18)) in a basal cytotoxicity assay with the rainbow trout gill cell line (RTgill-W1). Cells were exposed for 48 h in 96-well plates to increasing concentration of BACs in exposure medium containing 0, 60 μM bovine serum albumin (BSA) or 10% fetal bovine serum (FBS). Before and after exposure, BAC concentrations in exposure medium were analytically determined. Based on freely dissolved concentrations at the end of the exposure, median effect concentrations (EC(50)) decreased with increasing alkyl chain length up to 14 carbons. For BAC with alkyl chains of 12 or more carbons, EC(50)’s based on measured concentrations after exposure in supplement-free medium were up to 25-times lower than EC(50)’s calculated using nominal concentrations. When BSA or FBS was added to the medium, a decrease in cytotoxic potency of up to 22 times was observed for BAC with alkyl chains of eight or more carbons. The results of this study emphasize the importance of expressing the in vitro readouts as a function of a dose metric that is least influenced by assay setup to compare assay sensitivities and chemical potencies. |
| Databáze: | OpenAIRE |
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